IAJPS 2016, 3 (6), 582-585
A.Rajeshkanna et al
CODEN (USA): IAJPBB
ISSN 2349-7750
ISSN: 2349-7750
INDO AMERICAN JOURNAL OF
PHARMACEUTICAL SCIENCES Available online at: http://www.iajps.com
Research Article
ANTI CANCER ACTIVITY OF DATURA STRAMONIUM (FLOWERS) AGAINST HUMAN LIVER CANCER A. Rajeshkanna1*, D. Prabhakaran2, M. M. Senthamilselvi3, N. Muruganantham4, S. Solomon5 1
Department of Chemistry, Ananda College, Deva kottai, Tamil Nadu, India. 2 Chettinad Cement Corporation Ltd., Ariyalur, Tamil Nadu, India. 3 Principal,Government Arts College, Ariyalur, Tamil Nadu, India. 4 Department of Chemistry, Roever Engineering College, Perambalur, Tamil Nadu, India 5 Department of Chemistry, Periyar E.V.R.College (Autonomous), Trichy, Tamil Nadu, India.
Abstract: Our present investigation is focused on the anticancer activity of the compound isolated from the ethyl acetate fraction of flowers of Datura stramonium against human liver cancer HePG2 cell line by MTT assay using in-vitro method. The CTC50 value of the sample was 131.53 μg/ml against liver cancer HePG2 cell lines. Significant results were observed there by explaining the use of this plant in the traditional system of medicine. Keywords: MTT assay, anticancer activity, Datura stramonium, Liver cancer HePG2
Corresponding author: A.Rajeshkanna Assistant Professor Department of Chemistry Ananda College, Devakottai Sivagangai – 630303. Tamil Nadu, India. E.mail: rajchem1985@gmail.com Ph.No: 8531007125
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Please cite this article in press as A.Rajeshkanna et al, Anti Cancer Activity of Datura stramonium (flowers) Against Human Liver Cancer, Indo Am. J. P. Sci, 2016; 3(6).
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IAJPS 2016, 3 (6), 582-585
A.Rajeshkanna et al
INTRODUCTION: Medicinal plants have various effects on living systems which include sedative, analgesic, antipyretic, cardioprotective, antibacterial, antiviral and antiprotozoal among others [1]. The specific constituents which impact medicinal values on the plants can be derived from whole or parts of the plant such as stems, leaves, fruits, flowers, seeds and roots [2]. The growing public interest and awareness in herbal medicine have led the pharmaceutical industry and biomedical researchers to give more attention on medicinal plants [3]. Cancer is one of the ailments which cannot be completely subdued by chemotherapy. The chemotherapeutic agents though effective against various types of tumor, they are not totally free from side effects [4]. Datura stramonium (family: solanaceae) is a wildgrowing herb, known as Jimson weed. It also has several other names: thorn apple, angel’s trumpet, loco weed, etc. The incidence of D. stramonium poisoning is sporadic with a cluster of poisoning cases occurring mostly among adolescents. Some medicinal uses of the plant are its anti-inflammatory property of all part of the plants, stimulation of the central nervous system (CNS), respiratory decongestion, treatment of dental and skin infections
ISSN 2349-7750
and also in the treatment of toothache and alopecia [5-12]. MATERIALS AND METHODS: Collection of Flowers Fresh flowers of Datura stramonium were collected from S. Pudur, Sivagangai (Dt), Tamil Nadu, India, during the month of January and identified by Dr.S.John Britto, Director, The rapinat Herbarium and Centre for Molecular Systematics (Authentication No. AR001 dated: 08/01/2016). St.Joseph’s College (Campus), Trichy, Tamil Nadu, India. Extraction and fractionation Fresh flower (3 kg) of Datura stramonium collected at S. Pudur, Sivagangai (Dt), Tamil Nadu, India were extracted with 90% ethanol (5x500ml). The combined alcoholic extract was concentrated in vacuo and the aqueous extract was successively fractionated with petroleum ether (60-800C) (6x250ml), Peroxide free diethyl ether (4x250ml) and ethyl acetate (8x250ml). Petroleum ether fraction and diethyl ether fraction did not yield any isolable material. Ethyl acetate fraction on concentration yielded a dry powder which was dissolved in DMSO to get various concentrations and were used for further study.
MTT Assay method HePG2 cell line figures:
Fig: 1-5 Effect of the compound isolated from the ethyl acetate fraction of Datura stramonium flowers against human Liver cancer HePG2 Cell line in different concentrations
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IAJPS 2016, 3 (6), 582-585
A.Rajeshkanna et al
ISSN 2349-7750
Table 1: The CTC50 values of the compound isolated from the ethyl acetate fraction of Datura stramonium flowers against human Liver cancer HePG2 Cell line S. No 1 2 3 4 5
Concentration (µg/ml) 1000 500 250 125 62.5
% CTC50 Cytotoxicity (µg/ml) 79.43 64.15 58.63 49.52 37.48
CTC50
131.53
Graphical representation of the CTC50 values of the compound isolated from the ethyl acetate fraction of Datura stramonium flowers against human Liver cancer HePG2 Cell line. MTT Assay: MTT-Assay-Chemicals 3-(4,5–dimethyl thiazol–2–yl)–5–diphenyl tetrazolium bromide (MTT), Fetal Bovine serum (FBS), Phosphate Buffered Saline (PBS), Dulbecco’s Modified Eagle’s Medium (DMEM) and Trypsin were obtained from Sigma Aldrich Co, St Louis, USA. EDTA, Glucose and antibiotics from Hi-Media Laboratories Ltd., Mumbai. Dimethyl Sulfoxide (DMSO) and Propanol from E.Merck Ltd., Mumbai, India.
out in 96 microtitre plates (Tarsons India Pvt. Ltd., Kolkata, India).
Cell Lines and Culture Medium HePG2 (Liver cancer cell line) cell cultures were procured from National Centre for Cell Sciences (NCCS), Pune, India. Stock cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM). Medium was supplemented with 10% inactivated Fetal Bovine Serum (FBS), penicillin (100 IU/ml), streptomycin (100 g/ml) and amphotericin B (5 g/ml) in an humidified atmosphere of 5% CO2 at 37C until confluent. The cells were dissociated with TPVG solution (0.2% trypsin, 0.02% EDTA, 0.05% glucose in PBS). The stock cultures were grown in 25 cm2 culture flasks and all experiments were carried
Determination of Cell Viability by MTT Assays The monolayer cell culture was trypsinized and the cell count was adjusted to 1.0 x 105 cells/ml using medium containing 10% FBS and were used for the determination of cell viability by MTT assays as described by Francis and Rita (1986) respectively. The absorbance was measured using a microplate reader at a wavelength of 540 nm. The percentage growth inhibition was calculated using the following formula and concentration of test drug needed to inhibit cell growth by 50% (CTC50) values is generated from the dose-response curves for each cell line.
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Preparation of Test Solutions For cytotoxicity studies, each weighed test drugs were separately dissolved in distilled DMSO and volume was made up with DMEM supplemented with 2% inactivated FBS to obtain a stock solution of 1 mg/ml concentration and sterilized by filtration. Serially two fold dilutions were prepared from this for carrying out cytotoxic studies.
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IAJPS 2016, 3 (6), 582-585
A.Rajeshkanna et al
% Growth inhibition = Mean OD of individual test group 100 ×100 Mean OD of control group RESULT AND DISCUSSION: The different concentration of the compound isolated from the ethyl acetate fraction of Datura stramonium flowers were subjected for MTT assay and results are presented in table.1. The photographs (Fig. 1 to Fig. 5) show the effect of the compound on the human liver cancer HePG2 cell line. The sample concentrations of 1000μg/ml, 500 μg/ml, 250μg/ml, 125μg/ml and 62.5μg/ml show 79.43 μg/ml, 64.15 μg/ml, 58.63 μg/ml, 49.52 μg/ml, 37.48 μg/ml CTC50 value against the human liver cancer HePG2 cell line respectively. CONCLUSION: The MTT assay of the compound isolated from the ethyl acetate fraction of flowers of Datura stramonium shows that all concentrations are having anticancer activity. So, it could be concluded that the compound to have a high anticancer potential.
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8.Zagari A (1992). Medicinal plants Vol. 3, 5th ed. Tehran University Publication, No. 1810/3, Tehran, Iran. pp.889. 9.John, D. One hundred useful drugs of the Kani tribes of Trivandum forest divisions, Kerala, India.Inter. J. crude drug Res.1984; 22:17-39. 10.Darias, V., Brovo, L., Barquin, E., Horrera, D.M. and Fraile, C. Contribution to the ethnopharmcological study of the Canary Islands. J. Ethnopharml. 1986;18: 169-193. 11.De Foe V. and Senatore F . Medicinal plants and phytotherapy in the Amal Fitan Cost, Salerno province Campania, Southern Italy. J .Ethnopharml. 1993;39: 39-51. 12.Abebe, W. A survey of prescriptions used in traditional medicine in Gondar region, North West Ethiopia: general pharmaceutical practice. J. Ethnopharm. 1986;18: 147-165.
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